Bioprocessing Technology Institute, A*STAR-Agency for Science, Technology and Research, 20 Biopolis Way, #06-01 Centros, Singapore 138668, Singapore.
J Biotechnol. 2013 Sep 10;167(3):215-24. doi: 10.1016/j.jbiotec.2013.07.010. Epub 2013 Jul 19.
We report the first investigation of translational efficiency on a global scale, also known as translatome, of a Chinese hamster ovary (CHO) DG44 cell line producing monoclonal antibodies (mAb). The translatome data was generated via combined use of high resolution and streamlined polysome profiling technology and proprietary Nimblegen microarrays probing for more than 13K annotated CHO-specific genes. The distribution of ribosome loading during the exponential growth phase revealed the translational activity corresponding to the maximal growth rate, thus allowing us to identify stably and highly translated genes encoding heterogeneous nuclear ribonucleoproteins (Hnrnpc and Hnrnpa2b1), protein regulator of cytokinesis 1 (Prc1), glucose-6-phosphate dehydrogenase (G6pdh), UTP6 small subunit processome (Utp6) and RuvB-like protein 1 (Ruvbl1) as potential key players for cellular growth. Moreover, correlation analysis between transcriptome and translatome data sets showed that transcript level and translation efficiency were uncoupled for 95% of investigated genes, suggesting the implication of translational control mechanisms such as the mTOR pathway. Thus, the current translatome analysis platform offers new insights into gene expression in CHO cell cultures by bridging the gap between transcriptome and proteome data, which will enable researchers of the bioprocessing field to prioritize in high-potential candidate genes and to devise optimal strategies for cell engineering toward improving culture performance.
我们报告了首次对生产单克隆抗体的中国仓鼠卵巢（CHO）DG44 细胞系进行全球范围的翻译效率（也称为翻译组）研究。该翻译组数据是通过高分辨率和简化的多核糖体分析技术以及专有 Nimblegen 微阵列的组合使用生成的，该微阵列可探测超过 13000 个注释的 CHO 特异性基因。在指数增长阶段，核糖体的加载分布揭示了与最大生长速率相对应的翻译活性，从而使我们能够鉴定出稳定且高度翻译的基因，这些基因编码异质核核糖核蛋白（Hnrnpc 和 Hnrnpa2b1）、细胞分裂调控蛋白 1（Prc1）、葡萄糖-6-磷酸脱氢酶（G6pdh）、UTP6 小亚基加工体（Utp6）和 RuvB 样蛋白 1（Ruvbl1），它们可能是细胞生长的关键因素。此外，转录组和翻译组数据集之间的相关性分析表明，在 95%的研究基因中，转录水平和翻译效率是解耦的，这表明存在翻译控制机制，如 mTOR 途径。因此，当前的翻译组分析平台通过弥合转录组和蛋白质组数据之间的差距，为 CHO 细胞培养中的基因表达提供了新的见解，这将使生物处理领域的研究人员能够优先考虑高潜力的候选基因，并设计出优化的细胞工程策略，以提高培养性能。
